Ophthalmic compositions for treating ocular hypertension

ABSTRACT

This invention relates to potent potassium channel blocker compounds of Formula I or a formulation thereof for the treatment of glaucoma and other conditions which leads to elevated intraoccular pressure in the eye of a patient. This invention also relates to the use of such compounds to provide a neuroprotective effect to the eye of mammalian species, particularly humans.

[0001] This claims the benefit of U.S. Provisional Applications60/424,808 and 60/500,091.

BACKGROUND OF THE INVENTION

[0002] Glaucoma is a degenerative disease of the eye wherein theintraocular pressure is too high to permit normal eye function. As aresult, damage may occur to the optic nerve head and result inirreversible loss of visual function. If untreated, glaucoma mayeventually lead to blindness. Ocular hypertension, i.e., the conditionof elevated intraocular pressure without optic nerve head damage orcharacteristic glaucomatous visual field defects, is now believed by themajority of ophthalmologists to represent merely the earliest phase inthe onset of glaucoma.

[0003] There are several therapies for treating glaucoma and elevatedintraocular pressure, but the efficacy and the side effect profiles ofthese agents are not ideal. Recently potassium channel blockers werefound to reduce intraocular pressure in the eye and therefore provideyet one more approach to the treatment of ocular hypertension and thedegenerative ocular conditions related thereto. Blockage of potassiumchannels can diminish fluid secretion, and under some circumstances,increase smooth muscle contraction and would be expected to lower IOPand have neuroprotective effects in the eye. (see U.S. Pat. Nos.5,573,758 and 5,925,342; Moore, et al., Invest. Ophthalmol. Vis. Sci 38,1997; WO 89/10757, WO94/28900, and WO 96/33719).

SUMMARY OF THE INVENTION

[0004] This invention relates to the use of potent potassium channelblockers or a formulation thereof in the treatment of glaucoma and otherconditions which are related to elevated intraocular pressure in the eyeof a patient. This invention also relates to the use of such compoundsto provide a neuroprotective effect to the eye of mammalian species,particularly humans. More particularly this invention relates to thetreatment of glaucoma and/or ocular hypertension (elevated intraocularpressure) using novel indazole compounds having the structural formulaI:

Formula I

[0005] or a pharmaceutically acceptable salt, enantiomer, diastereomeror mixture thereof:

[0006] wherein,

[0007] R represents hydrogen, or C₁₋₆ alkyl;

[0008] X represents —(CHR₇)_(p)—, —(CHR₇)_(p)CO—;

[0009] Y represents —CO(CH₂)_(n)—, CH₂, or —CH(OR)—;

[0010] Q represents CR^(y);

[0011] R^(y) represents H, or C₁₋₆ alkyl;

[0012] R_(w) represents H, C₁₋₆ alkyl, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆alkyl, —SO₂N(R)₂, —SO₂C₁₋₆ alkyl, —SO₂C₆₋₁₀ aryl, NO₂, CN or —C(O)N(R)₂;

[0013] R₂ represents hydrogen, C₁₋₁₀ alkyl, OH, C₂₋₆ alkenyl, C₁₋₆alkylSR, —(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆ alkoxy, —(CH₂)_(n)C₃₋₈cycloalkyl, —(CH₂)_(n)C₃₋₁₀ heterocyclyl, —N(R)₂, —COOR, or—(CH₂)_(n)C₆₋₁₀ aryl, said alkyl, heterocyclyl, or aryl optionallysubstituted with 1-3 groups selected from R^(a);

[0014] R₃ represents hydrogen, C₁₋₁₀ alkyl, —(CH₂)_(n)C₃₋₈ cycloalkyl,—(CH₂)_(n)C₃₋₁₀ heterocyclyl, —(CH₂)_(n)COOR, —(CH₂)_(n)C₆₋₁₀ aryl,—(CH₂)_(n)NHR₈, —(CH₂)_(n)N(R)₂, —(CH₂)_(n)N(R₈)₂, —(CH₂)_(n)NHCOOR,—(CH₂)_(n)N(R₈)CO₂R, —(CH₂)_(n)N(R₈)COR, —(CH₂)_(n)NHCOR,—(CH₂)_(n)CONH(R₈), aryl, —(CH₂)_(n)C₁₋₆ alkoxy, CF₃, —(CH₂)_(n)SO₂R,—(CH₂)_(n)SO₂N(R)₂, —(CH₂)_(n)CON(R)₂, —(CH₂)_(n)CONHC(R)₃,—(CH₂)_(n)CONHC(R)₂CO₂R, —(CH₂)_(n)COR₈, nitro, cyano or halogen, saidalkyl, alkoxy, heterocyclyl, or aryl optionally substituted with 1-3groups of R^(a);

[0015] or R₂ and R₃ taken together with the intervening Q form a 3-10membered carbocyclic or heterocyclic carbon ring optionally interruptedby 1-2 atoms of O, S, C(O) or NR, and optionally having 1-4 doublebonds, and optionally substituted by 1-3 groups selected from R^(a);

[0016] or R₂ and R₃ taken together with the intervening Q represent OR;

[0017] R₄ and R₅ independently represent hydrogen, C₁₋₆ alkoxy, OH, C₁₋₆alkyl, COOR, SO_(q)C₁₋₆ alkyl, COC₁₋₆ alkyl, SO₃H, —O(CH₂)_(n)N(R)₂,—O(CH₂)_(n)CO₂R, —OPO(OH)₂, CF₃, OCF₃—N(R)₂, nitro, cyano, C₁₋₆alkylamino, or halogen; and

[0018] R₆ represents hydrogen, C₁₋₁₀ alkyl, —(CH₂)_(n)C₆₋₁₀ aryl,NR_(c)R_(d), —NR(CH₂)_(n)C₆₋₁₀ aryl, —N((CH₂)_(n)C₆₋₁₀ aryl)₂,—(CH₂)_(n)C₃₋₁₀ heterocyclyl, —NR(CH₂)_(n)C₃₋₁₀ heterocyclyl,—N((CH₂)_(n)C₃₋₁₀ heterocyclyl)₂ (C₆₋₁₀ aryl)O—, —(CH₂)_(n)C₃₋₈cycloalkyl, —COOR, —C(O)CO₂R, said aryl, heterocyclyl and alkyloptionally substituted with 1-3 groups selected from R^(a), wherein theR^(a)(s) can be attached to any carbon atom or heteroatom selected fromN and S;

[0019] R_(c) and R_(d) independently represent H, C₁₋₆ alkyl, C₂₋₆alkenyl, C₁₋₆ alkylSR, —(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆ alkoxy, or—(CH₂)_(n)C₃₋₈ cycloalkyl;

[0020] or R_(c) and R_(d) taken together with the intervening N atomform a 4-10 membered heterocyclic carbon ring optionally interrupted by1-2 atoms of O, S, C(O) or NR, and optionally having 1-4 double bonds,and optionally substituted by 1-3 groups selected from R^(a);

[0021] R₇ represents hydrogen, C₁₋₆ alkyl, —(CH₂)_(n)COOR or—(CH₂)_(n)N(R)₂,

[0022] R₈ represents —(CH₂)_(n)C₃₋₈ cycloalkyl, —(CH₂)_(n 3-10)heterocyclyl, C₁₋₆ alkoxy or —(CH₂)_(n)C₅₋₁₀ heteroaryl, —(CH₂)_(n)C₆₋₁₀aryl said heterocyclyl, aryl or heteroaryl optionally substituted with1-3 groups selected from R^(a);

[0023] Ra represents F, Cl, Br, I, CF₃, N(R)₂, NO₂, CN, —O—, —COR₈,—CONHR₈, —CON(R₈)₂, —O(CH₂)_(n)COOR, —NH(CH₂)_(n)OR, —COOR, —OCF₃,CF₂CH₂OR, —NHCOR, —SO₂R, —SO₂NR₂, —SR, (C₁-C₆ alkyl)O—,—(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆ alkoxy, (aryl)O—, —(CH₂)_(n)OH,(C₁-C₆ alkyl)S(O)M—, H₂N—C(NH)—, (C₁-C₆ alkyl)C(O)—, (C₁-C₆alkyl)OC(O)NH—, —(C₁-C₆ alkyl)NR_(w)(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w),—(C₁-C₆ alkyl)O(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₁-C₆alkyl)S(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₁-C₆ alkyl)—C₃₋₁₀heterocyclyl-R_(w), —(CH₂)_(n)—Z¹—C(═Z²)N(R)₂, —(C₂₋₆alkenyl)NR_(w)(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆alkenyl)O(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆alkenyl)S(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆ alkenyl)-C₃₋₁₀heterocyclyl-R_(w), —(C₂₋₆ alkenyl)—Z¹—C(═Z²)N(R)₂, —(CH₂)_(n)SO₂R,—(CH₂)_(n)SO₃H, —(CH₂)_(n)PO(OR)₂, C₃₋₁₀cycloalkyl, C₆₋₁₀ aryl, C₃₋₁₀heterocyclyl, C₂₋₆ alkenyl, and C₁-C₁₀ alkyl, said alkyl, alkenyl,alkoxy, heterocyclyl and aryl optionally substituted with 1-3 groupsselected from C₁-C₆ alkyl, halogen, (CH₂)_(n)OH, CN, NO₂, CON(R)₂ andCOOR;

[0024] Z¹ and Z² independently represents NR_(w), O, CH₂, or S;

[0025] m is 0-3;

[0026] n is 0-3;

[0027] p is 0-3 and

[0028] q is 0-2.

[0029] This and other aspects of the invention will be realized uponinspection of the invention as a whole.

DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention is directed to novel potassium channelblockers of Formula I. It also relates to a method for decreasingelevated intraocular pressure or treating glaucoma by administration,preferably topical or intra-camaral administration, of a compositioncontaining a potassium channel blocker of Formula I describedhereinabove and a pharmaceutically acceptable carrier.

[0031] One embodiment of this invention is realized when p is 1-3.

[0032] Another embodiment of this invention is realized when Y is—CO(CH₂)_(n) and all other variables are as originally described. Asubembodiment of this invention is realized when n is 0.

[0033] Another embodiment of this invention is realized when Y is CH(OR)and all other variables are as originally described.

[0034] In another embodiment R_(w) is selected from H, C₁₋₆ alkyl,—C(O)C₁₋₆ alkyl and C(O)N(R)₂ and all other variables are as originallydescribed.

[0035] In another embodiment X is —(CHR₇)_(p)—, p is 1-3 and all othervariables are as originally described.

[0036] In another embodiment X is —(CHR₇)_(p)CO—, p is 1-3 and all othervariables are as originally described.

[0037] Still another embodiment of this invention is realized when R₆ isC₁₋₁₀ alkyl, (CH₂)_(n)C₆₋₁₀ aryl, (CH₂)_(n)C₃₋₁₀ heterocyclyl,NR_(c)R_(d) or (CH₂)_(n)C₃₋₈ cycloalkyl, said alkyl, aryl, heterocyclyland cycloalkyl optionally substituted with 1 to 3 groups of R^(a), andall other variables are as originally described.

[0038] Yet another embodiment of this invention is realized when R₆ isC₁₋₁₀ alkyl, (CH₂)_(n)C₆₋₁₀ aryl, or (CH₂)_(n)C₃₋₁₀ heterocyclyl, saidalkyl, aryl, and heterocyclyl optionally substituted with 1 to 3 groupsof R^(a), and all other variables are as originally described.

[0039] Yet another embodiment of this invention is realized when R₇ ishydrogen or C₁₋₆ alkyl, and all other variables are as originallydescribed.

[0040] Yet another embodiment of this invention is realized when Y is—CO(CH₂)_(n), and n is 0.

[0041] Still another embodiment of this invention is realized when Y is—CO(CH₂)_(n), R₂ is C₁₋₁₀ alkyl or C₁₋₆ alkylOR and R₃ is C₁₋₁₀ alkyl,(CH₂)_(n)C₃₋₁₀ heterocyclyl, X is —(CHR₇)_(p)CO—, and p is 1-3 saidheterocyclyl and alkyl optionally substituted with 1 to 3 groups ofR^(a). A subembodiment of this invention is realized when n is 0.

[0042] Another embodiment of the instant invention is realized whenR^(a) is selected from F, Cl, Br, I, CF₃, N(R)₂, NO₂, CN, —O—, —CONHR₈,—CON(R₈)₂, —O(CH₂)_(n)COOR, —NH(CH₂)_(n)OR, —COOR, —OCF₃, —NHCOR, —SO₂R,—SO₂NR₂, —SR, (C₁-C₆ alkyl)O—, —(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆alkoxy, (aryl)O—, —OH, (C₁-C₆ alkyl)S(O)_(m)—, H₂N—C(NH)—, (C₁-C₆alkyl)C(O)—, (C₁-C₆ alkyl)OC(O)NH—, —(C₁-C₆ alkyl)NR_(w)(CH₂)_(n)C₃₋₁₀heterocyclyl R_(w), —(CH₂)_(n)—Z¹—C(═Z²)N(R)₂, —(C₂₋₆alkenyl)NR_(w)(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆alkenyl)—Z¹—C(═Z²)N(R)₂, —(CH₂)_(n)SO₂R, —(CH₂)_(n)SO₃H,—(CH₂)_(n)PO(OR)₂, C₂₋₆ alkenyl, and C₁-C₁₀ alkyl, said alkyl andalkenyl, optionally substituted with 1-3 groups selected from C₁-C₆alkyl, and COOR;

[0043] Examples of compounds to be used in this invention are found inTables 1 and 2: TABLE 1

R1 R2

[0044] TABLE 2

wherein R₁ is

and X is N or CH;

[0045] or a pharmaceutically acceptable salt, enantiomer, diastereomeror mixture thereof.

[0046] The invention is described herein in detail using the termsdefined below unless otherwise specified.

[0047] The compounds of the present invention may have asymmetriccenters, chiral axes and chiral planes, and occur as racemates, racemicmixtures, and as individual diastereomers, with all possible isomers,including optical isomers, being included in the present invention. (SeeE. L. Eliel and S. H. Wilen Stereochemistry of Carbon Compounds (JohnWiley and Sons, New York 1994), in particular pages 1119-1190)

[0048] When any variable (e.g. aryl, heterocycle, R¹, R⁶ etc.) occursmore than one time in any constituent, its definition on each occurrenceis independent at every other occurrence. Also, combinations ofsubstituents/or variables are permissible only if such combinationsresult in stable compounds.

[0049] When R^(a) is —O— and attached to a carbon it is referred to as acarbonyl group and when it is attached to a nitrogen (e.g., nitrogenatom on a pyridyl group) or sulfur atom it is referred to a N-oxide andsulfoxide group, respectively.

[0050] The term “alkyl” refers to a monovalent alkane (hydrocarbon)derived radical containing from 1 to 10 carbon atoms unless otherwisedefined. It may be straight, branched or cyclic. Preferred alkyl groupsinclude methyl, ethyl, propyl, isopropyl, butyl, t-butyl, cyclopropylcyclopentyl and cyclohexyl. When the alkyl group is said to besubstituted with an alkyl group, this is used interchangeably with“branched alkyl group”.

[0051] Cycloalkyl is a specie of alkyl containing from 3 to 15 carbonatoms, unless otherwise defined, without alternating or resonatingdouble bonds between carbon atoms. It may contain from 1 to 4 rings,which are fused. Examples of such cycloalkyl elements include, but arenot limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl andcycloheptyl.

[0052] Alkenyl is C₂-C₆ alkenyl.

[0053] Alkoxy refers to an alkyl group of indicated number of carbonatoms attached through an oxygen bridge, with the alkyl group optionallysubstituted as described herein. Said groups are those groups of thedesignated length in either a straight or branched configuration and iftwo or more carbon atoms in length, they may include a double or atriple bond. Exemplary of such alkoxy groups are methoxy, ethoxy,propoxy, isopropoxy, butoxy, isobutoxy, tertiary butoxy, pentoxy,isopentoxy, hexoxy, isohexoxy allyloxy, propargyloxy, and the like.

[0054] Halogen (halo) refers to chlorine, fluorine, iodine or bromine.

[0055] Aryl refers to aromatic rings e.g., phenyl, substituted phenyland the like, as well as rings which are fused, e.g., naphthyl,phenanthrenyl and the like. An aryl group thus contains at least onering having at least 6 atoms, with up to five such rings being present,containing up to 22 atoms therein, with alternating (resonating) doublebonds between adjacent carbon atoms or suitable heteroatoms. Examples ofaryl groups are phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl,phenanthryl, anthryl or acenaphthyl and phenanthrenyl, preferablyphenyl, naphthyl or phenanthrenyl. Aryl groups may likewise besubstituted as defined. Preferred substituted aryls include phenyl andnaphthyl.

[0056] The term heterocyclyl or heterocyclic, as used herein, representsa stable 3- to 7-membered monocyclic or stable 8- to 11-memberedbicyclic heterocyclic ring which is either saturated or unsaturated, andwhich consists of carbon atoms and from one to four heteroatoms selectedfrom the group consisting of N, O, and S, and including any bicyclicgroup in which any of the above-defined heterocyclic rings is fused to abenzene ring. The heterocyclic ring may be attached at any heteroatom orcarbon atom which results in the creation of a stable structure. A fusedheterocyclic ring system may include carbocyclic rings and need includeonly one heterocyclic ring. The term heterocycle or heterocyclicincludes heteroaryl moieties. Examples of such heterocyclic elementsinclude, but are not limited to, azepinyl, benzimidazolyl,benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl,benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl,cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl,dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone,dihydropyrrolyl, 1,3-dioxolanyl, furyl, imidazolidinyl, imidazolinyl,imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl,isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl,morpholinyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl,2-oxopiperazinyl, 2-oxopiperdinyl, 2-oxopyrrolidinyl, piperidyl,piperazinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl, pyridazinyl,pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl,quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl,tetrahydroquinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide,thiazolyl, thiazolinyl, thienofuryl, thienothienyl, and thienyl.Preferably, heterocycle is selected from 2-azepinonyl, benzimidazolyl,2-diazapinonyl, dihydroimidazolyl, dihydropyrrolyl, imidazolyl,2-imidazolidinonyl, indolyl, isoquinolinyl, morpholinyl, piperidyl,piperazinyl, pyridyl, pyrrolidinyl, 2-piperidinonyl, 2-pyrimidinonyl,2-pyrollidinonyl, quinolinyl, tetrahydrofuryl, tetrahydroisoquinolinyl,and thienyl.

[0057] The term “heteroatom” means O, S or N, selected on an independentbasis.

[0058] The term “heteroaryl” refers to a monocyclic aromatic hydrocarbongroup having 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to10 atoms, containing at least one heteroatom, O, S or N, in which acarbon or nitrogen atom is the point of attachment, and in which one ortwo additional carbon atoms is optionally replaced by a heteroatomselected from O or S, and in which from 1 to 3 additional carbon atomsare optionally replaced by nitrogen heteroatoms, said heteroaryl groupbeing optionally substituted as described herein. Examples of suchheterocyclic elements include, but are not limited to, benzimidazolyl,benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl,benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl,cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl,dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, furyl,imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl,isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, pyridyl,pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl,quinolinyl, quinoxalinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,thiazolyl, thienofuryl, thienothienyl, thienyl and triazolyl. Additionalnitrogen atoms may be present together with the first nitrogen andoxygen or sulfur, giving, e.g., thiadiazole.

[0059] This invention is also concerned with compositions and methods oftreating ocular hypertension or glaucoma by administering to a patientin need thereof one of the compounds of formula I in combination with aβ-adrenergic blocking agent such as timolol, betaxolol, levobetaxolol,carteolol, levobunolol, a parasympathomimetic agent such as epinephrine,iopidine, brimonidine, clonidine, para-aminoclonidine, carbonicanhydrase inhibitor such as dorzolamide, acetazolamide, metazolamide orbrinzolamide, an EP4 agonist (such as those disclosed in WO 02/24647, WO02/42268, EP 1114816, WO 01/46140 and WO 01/72268), a prostaglandin suchas latanoprost, travaprost, unoprostone, rescula, S1033 (compounds setforth in U.S. Pat. Nos. 5,889,052; 5,296,504; 5,422,368; and 5,151,444);a hypotensive lipid such as lumigan and the compounds set forth in U.S.Pat. No. 5,352,708; a neuroprotectant disclosed in U.S. Pat. No.4,690,931, particularly eliprodil and R-eliprodil as set forth in WO94/13275, including memantine; or an agonist of 5-HT2 receptors as setforth in PCT/U.S.OO/31247, particularly1-(2-aminopropyl)-3-methyl-1H-imdazol-6-ol fumarate and2-(3-chloro-6-methoxy-indazol-1-yl)-1-methyl-ethylamine. An example of ahypotensive lipid (the carboxylic acid group on the α-chain link of thebasic prostaglandin structure is replaced with electrochemically neutralsubstituents) is that in which the carboxylic acid group is replacedwith a C₁₋₆ alkoxy group such as OCH₃ (PGF_(2a) 1-OCH₃), or a hydroxygroup (PGF_(2a) 1-OH).

[0060] Preferred potassium channel blockers are calcium activatedpotassium channel blockers. More preferred potassium channel blockersare high conductance, calcium activated potassium (Maxi-K) channelblockers. Maxi-K channels are a family of ion channels that areprevalent in neuronal, smooth muscle and epithelial tissues and whichare gated by membrane potential and intracellular Ca²⁺.

[0061] The present invention is based upon the finding that maxi-Kchannels, if blocked, inhibit aqueous humor production by inhibiting netsolute and H₂O efflux and therefore lower IOP. This finding suggeststhat maxi-K channel blockers are useful for treating otherophthamological dysfunctions such as macular edema and maculardegeneration. It is known that lowering IOP promotes blood flow to theretina and optic nerve. Accordingly, the compounds of this invention areuseful for treating macular edema and/or macular degeneration.

[0062] It is believed that maxi-K channel blockers which lower IOP areuseful for providing a neuroprotective effect. They are also believed tobe effective for increasing retinal and optic nerve head blood velocityand increasing retinal and optic nerve oxygen by lowering IOP, whichwhen coupled together benefits optic nerve health. As a result, thisinvention further relates to a method for increasing retinal and opticnerve head blood velocity, increasing retinal and optic nerve oxygentension as well as providing a neuroprotective effect or a combinationthereof.

[0063] A number of marketed drugs function as potassium channelantagonists. The most important of these include the compoundsGlyburide, Glipizide and Tolbutamide. These potassium channelantagonists are useful as antidiabetic agents. The compounds of thisinvention may be combined with one or more of these compounds to treatdiabetes.

[0064] Potassium channel antagonists are also utilized as Class 3antiarrhythmic agents and to treat acute infarctions in humans. A numberof naturally occuring toxins are known to block potassium channelsincluding Apamin, Iberiotoxin, Charybdotoxin, Noxiustoxin, Kaliotoxin,Dendrotoxin(s), mast cell degranuating (MCD) peptide, and β-Bungarotoxin(β-BTX). The compounds of this invention may be combined with one ormore of these compounds to treat arrhythmias.

[0065] Depression is related to a decrease in neurotransmitter release.Current treatments of depression include blockers of neurotransmitteruptake, and inhibitors of enzymes involved in neurotransmitterdegradation which act to prolong the lifetime of neurotransmitters.

[0066] Alzheimer's disease is also characterized by a diminishedneurotransmitter release. Three classes of drugs are being investigatedfor the treatment of Alzheimer's disease cholinergic potentiators suchas the anticholinesterase drugs (e.g., physostigmine (eserine), andTacrine (tetrahydroaminocridine)); nootropics that affect neuronmetabolism with little effect elsewhere (e.g., Piracetam, Oxiracetam;and those drugs that affect brain vasculature such as a mixture ofergoloid mesylates amd calcium channel blocking drugs includingNimodipine. Selegiline, a monoamine oxidase B inhibitor which increasesbrain dopamine and norepinephrine has reportedly caused mild improvementin some Alzheimer's patients. Aluminum chelating agents have been ofinterest to those who believe Alzheimer's disease is due to aluminumtoxicity. Drugs that affect behavior, including neuroleptics, andanxiolytics have been employed. Anxiolytics, which are mildtranquilizers, are less effective than neuroleptics The presentinvention is related to novel compounds which are useful as potassiumchannel antagonists.

[0067] The compounds within the scope of the present invention exhibitpotassium channel antagonist activity and thus are useful in disordersassociated with potassium channel malfunction. A number of cognitivedisorders such as Alzheimer's Disease, memory loss or depression maybenefit from enhanced release of neurotransmitters such as serotonin,dopamine or acetylcholine and the like. Blockage of Maxi-K channelsmaintains cellular depolarization and therefore enhances secretion ofthese vital neurotransmitters.

[0068] The compounds of this invention may be combined withanticholinesterase drugs such as physostigmine (eserine) and Tacrine(tetrahydroaminocridine), nootropics such as Piracetam, Oxiracetam,ergoloid mesylates, selective calcium channel blockers such asNimodipine, or monoamine oxidase B inhibitors such as Selegiline, in thetreatment of Alzheimer's disease. The compounds of this invention mayalso be combined with Apamin, Iberiotoxin, Charybdotoxin, Noxiustoxin,Kaliotoxin, Dendrotoxin(s), mast cell degranuating (MCD) peptide,β-Bungarotoxin (β-BTX) or a combination thereof in treating arrythmias.The compounds of this invention may further be combined with Glyburide,Glipizide, Tolbutamide or a combination thereof to treat diabetes.

[0069] The herein examples illustrate but do not limit the claimedinvention. Each of the claimed compounds are potassium channelantagonists and are thus useful in the described neurological disordersin which it is desirable to maintain the cell in a depolarized state toachieve maximal neurotransmitter release. The compounds produced in thepresent invention are readily combined with suitable and knownpharmaceutically acceptable excipients to produce compositions which maybe administered to mammals, including humans, to achieve effectivepotassium channel blockage.

[0070] For use in medicine, the salts of the compounds of formula I willbe pharmaceutically acceptable salts. Other salts may, however, beuseful in the preparation of the compounds according to the invention orof their pharmaceutically acceptable salts. When the compound of thepresent invention is acidic, suitable “pharmaceutically acceptablesalts” refers to salts prepared form pharmaceutically acceptablenon-toxic bases including inorganic bases and organic bases. Saltsderived from inorganic bases include aluminum, ammonium, calcium,copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous,potassium, sodium, zinc and the like. Particularly preferred are theammonium, calcium, magnesium, potassium and sodium salts. Salts derivedfrom pharmaceutically acceptable organic non-toxic bases include saltsof primary, secondary and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, such as arginine, betaine caffeine, choline,N,N¹-dibenzylethylenediamine, diethylamin, 2-diethylaminoethanol,2-dimethylaminoethanol, ethanolamine, ethylenediamine,N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine,hydrabamine, isopropylamine, lysine, methylglucamine, morpholine,piperazine, piperidine, polyamine resins, procaine, purines,theobromine, triethylamine, trimethylamine tripropylamine, tromethamineand the like.

[0071] When the compound of the present invention is basic, salts may beprepared from pharmaceutically acceptable non-toxic acids, includinginorganic and organic acids. Such acids include acetic, benzenesulfonic,benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic,glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and thelike. Particularly preferred are citric, hydrobromic, hydrochloric,maleic, phosphoric, sulfuric and tartaric acids.

[0072] The preparation of the pharmaceutically acceptable saltsdescribed above and other typical pharmaceutically acceptable salts ismore fully described by Berg et al., “Pharmaceutical Salts,” J. Pharm.Sci., 1977:66:1-19.

[0073] As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specific amounts, aswell as any product which results, directly or indirectly, fromcombination of the specific ingredients in the specified amounts.

[0074] When a compound according to this invention is administered intoa human subject, the daily dosage will normally be determined by theprescribing physician with the dosage generally varying according to theage, weight, sex and response of the individual patient, as well as theseverity of the patient's symptoms.

[0075] The maxi-K channel blockers used can be administered in atherapeutically effective amount intravaneously, subcutaneously,topically, transdermally, parenterally or any other method known tothose skilled in the art. Ophthalmic pharmaceutical compositions arepreferably adapted for topical administration to the eye in the form ofsolutions, suspensions, ointments, creams or as a solid insert.Ophthalmic formulations of this compound may contain from 0.01 ppm to 1%and especially 0.1 ppm to 1% of medicament. Higher dosages as, forexample, about 10% or lower dosages can be employed provided the dose iseffective in reducing intraocular pressure, treating glaucoma,increasing blood flow velocity or oxygen tension. For a single dose,from between 0.01 to 5000 ng, preferably 0.1 to 500 ng, and especially 1to 100 ng of the compound can be applied to the human eye.

[0076] The pharmaceutical preparation which contains the compound may beconveniently admixed with a non-toxic pharmaceutical organic carrier, orwith a non-toxic pharmaceutical inorganic carrier. Typical ofpharmaceutically acceptable carriers are, for example, water, mixturesof water and water-miscible solvents such as lower alkanols oraralkanols, vegetable oils, polyalkylene glycols, petroleum based jelly,ethyl cellulose, ethyl oleate, carboxymethyl-cellulose,polyvinylpyrrolidone, isopropyl myristate and other conventionallyemployed acceptable carriers. The pharmaceutical preparation may alsocontain non-toxic auxiliary substances such as emulsifying, preserving,wetting agents, bodying agents and the like, as for example,polyethylene glycols 200, 300, 400 and 600, carbowaxes 1,000, 1,500,4,000, 6,000 and 10,000, antibacterial components such as quaternaryammonium compounds, phenylmercuric salts known to have cold sterilizingproperties and which are non-injurious in use, thimerosal, methyl andpropyl paraben, benzyl alcohol, phenyl ethanol, buffering ingredientssuch as sodium borate, sodium acetates, gluconate buffers, and otherconventional ingredients such as sorbitan monolaurate, triethanolamine,oleate, polyoxyethylene sorbitan monopalmitylate, dioctyl sodiumsulfosuccinate, monothioglycerol, thiosorbitol, ethylenediaminetetracetic acid, and the like. Additionally, suitable ophthalmicvehicles can be used as carrier media for the present purpose includingconventional phosphate buffer vehicle systems, isotonic boric acidvehicles, isotonic sodium chloride vehicles, isotonic sodium boratevehicles and the like. The pharmaceutical preparation may also be in theform of a microparticle formulation. The pharmaceutical preparation mayalso be in the form of a solid insert. For example, one may use a solidwater soluble polymer as the carrier for the medicament. The polymerused to form the insert may be any water soluble non-toxic polymer, forexample, cellulose derivatives such as methylcellulose, sodiumcarboxymethyl cellulose, (hydroxyloweralkyl cellulose), hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose;acrylates such as polyacrylic acid salts, ethylacrylates,polyactylamides; natural products such as gelatin, alginates, pectins,tragacanth, karaya, chondrus, agar, acacia; the starch derivatives suchas starch acetate, hydroxymethyl starch ethers, hydroxypropyl starch, aswell as other synthetic derivatives such as polyvinyl alcohol, polyvinylpyrrolidone, polyvinyl methyl ether, polyethylene oxide, neutralizedcarbopol and xanthan gum, gellan gum, and mixtures of said polymer.

[0077] Suitable subjects for the administration of the formulation ofthe present invention include primates, man and other animals,particularly man and domesticated animals such as cats and dogs.

[0078] The pharmaceutical preparation may contain non-toxic auxiliarysubstances such as antibacterial components which are non-injurious inuse, for example, thimerosal, benzalkonium chloride, methyl and propylparaben, benzyldodecinium bromide, benzyl alcohol, or phenylethanol;buffering ingredients such as sodium chloride, sodium borate, sodiumacetate, sodium citrate, or gluconate buffers; and other conventionalingredients such as sorbitan monolaurate, triethanolamine,polyoxyethylene sorbitan monopalmitylate, ethylenediamine tetraaceticacid, and the like.

[0079] The ophthalmic solution or suspension may be administered asoften as necessary to maintain an acceptable IOP level in the eye. It iscontemplated that administration to the mamalian eye will be about onceor twice daily.

[0080] For topical ocular administration the novel formulations of thisinvention may take the form of solutions, gels, ointments, suspensionsor solid inserts, formulated so that a unit dosage comprises atherapeutically effective amount of the active component or somemultiple thereof in the case of a combination therapy.

[0081] The following examples given by way of illustration isdemonstrative of the present invention.

[0082] Definitions of the terms used in the examples are as follows:

[0083] SM—Starting material,

[0084] DMSO—dimethyl sulfoxide,

[0085] TLC—thin layer chromatography,

[0086] SGC—silica gel chromatography,

[0087] PhMgBr—phenylmagnesiumbromide

[0088] h=hr=hour,

[0089] THF—tetrahydrofuran,

[0090] DMF—dimethylformamide,

[0091] min—minute,

[0092] LC/MS—liquid chromatography/mass spectrometry,

[0093] HPLC—high performance liquid chromatography,

[0094] PyBOP—Benzotriazol-1-yloxytris-(dimethyl amino)phosphoniumhexafluorophosphate,

[0095] equiv=eq=equivalent,

[0096] NBS—N-Bromosuccinamide and

[0097] AIBN—2,2′-azobisisobutyronitrile.

[0098] The compounds of this invention can be made, with modificationwhere appropriate, in accordance with Schemes 1 through 4. Examples 1-3are also produced in accordance with Schemes 1 and/or 2.

[0099] In Schemes 1 and 2 nitroanisole is brominated using NBS, AIBN andbenzoyl peroxide. Treatment of the bromonitroanisole with potassiumcyanide yielded the cyanonitroanisole. Conversion of the nitro group toan amine is accomplished by hydrogenation. The amine is then treatedwith sodium nitrite and HCl to yield the indazole ring. In this reactionas soon as the diazonium is generated by nitrosation of the anilinemoiety it is trapped intramolecularily by the acidic benzyl cyanide.Tautomerization of the resultant derivative gives the indazole nucleus.Treatment of the nitrite with a Gringard followed by hydrolysis of theresultant imino-magnesium complex gives the desired alkyl/aryl ketone.

Preparative Example 1

[0100]

[0101] In a 500 mL flask was charged 336 mmoles (13.44 g; 60%) of NaH.Under argon 150 mL of DMSO was added, followed by dropwise addition of32 mL of ethyl cyanoacetate (2.2 equiv.; 352 mmloes) at 5° C. After allthe addition the reaction was warmed upto room temperature over 1 h. 30g of starting nitro benzene derivative was added (160 mmoles) as apowder. The reaction mixture was heated in a closed system at 90° C. for8 hours. Acidification and standard work-up gave a crude oily residuewhich was purified over a silica-gel column to give 39 g of desiredcrystalline product which was decarboxylated to give the benzyl nitrileas follows. Thirty eight grams of SM obtained above was dissolved in 400mL of 1N sodium carbonate. The homogenous solution was stirred at rt fortwo days. TLC analysis indicated competion of reaction. The reactionmixture was acidified and extratced with ethyl acetate (100 mL×4). Thecombined organic phases was dried over sodium sulphate and concentratedand residue was subjected to SGC to give the desired product.

[0102] 1H NMR CDCL3: 7.72 (1H, d, J=3 Hz); 7.61 (1H, d, J=8.5 Hz); 7.25(1H, dd, J=3 and 8.5 Hz); 4.17 (2H, s); 3.94 (3H, s). LCMS [M+H]=193.

Preparative Example 2

[0103]

[0104] 10 g of benzylnitrile derivative was dissolved in THF 20 mLfollowed by dilution with 50 mL of methanol. The reaction mixture wastaken in a pressure tube, Pd-C (10% wt/10 mole %) was added and thereaction mixture was hydrogenated at 40 psi. After the requisite amountof hydrogen for the reduction of the NO₂ group was consumed the reactionwas stopped. TLC analysis indicated a spot to spot conversion. Thereaction mixture was filtered over a pad of celite and the filtrate wasconcentrated to a solid and used in the next step directly. Crudeaniline derivative (52 mmoles was dissloved/suspended in 2N HCl (150mL), cooled to 5° C. followed by the addition of 5.4 g of sodium nitritein 10 mL of water. The reaction mixture was allowed to stir for 1 h withgradual warming to room temperature. TLC analysis indicated completeconsumption of SM and the formation of a new spot. The reaction mixturewas extratced with ethyl acetate (100 mL×4); organic phase wascollected, dried and concentrated. The residue was purified by SGC togive desired product. LCMS [M+H]=174

Preparative Example 3

[0105]

[0106] Nitrile (1.5 g) obtained from Preparative Example 2 was dissolvedin 20 mL of dry THF and under argon 3 equiv. of PhMgBr (1M in THF) wasadded at 5° C. The reaction mixture was stirred at room temperature for1 h. The reaction was carefully quenched by addition of water and 1N HCl(15 mL). The quenched reaction mixture was stirred at room temperaturefor 1 hour then extracted with ethyl acetate (20 mL×3); combined organicphases were dried over sodium sulfate and concentrated to a solidresidue which was azeotroped with toluene three times. LCMS [M+H]=253

Preparative Example 4

[0107]

[0108] Weighed out 4.15 g of indazole and azeoptroped water with 2toluene (100 ml) washings, pulling off toluene azeotrope by rotovap.Dried thoroughly under high vaccuum and performed argon purges.Dissolved in 40 ml dry THF and 92 ml dry ether under argon. Cooled to 5°C. in ice water bath. Charged 3 eq of isopropylmagnesium chloride ((6 mlof a 2M solution in THF) and stired for 0.5 hr at room temp. Carefullycharged 1N HCl (240 ml) and stired for 1 h. Monitored reaction by TLC.Extracted with EtOAc, rotovaped and produced desired product. LCMS[M+H]=219

Preparative Example 5

[0109]

[0110] Step A:

[0111] 100 g of 2-fluoro-4-methoxy-acetophenone in 400 mL of ethyleneglycol was stirred at room temperature with hydrazine (0.624 mol, 20 g)for 4 h after which the reaction mixture was heated to 150° C. for 48 h.TLC analysis indicated complete reaction. Partitioned the reactionmixture into dichloromethane and brine. Dried organic phase over sodiumsulphate and evaporated to a solid. Re-crystallized fromhexane/dicholomethane gave 6-methoxy-3-methyl-1H-indazole.

[0112] 1H NMR (CDCL3): 7.5 (1H, d, 7.5 Hz); 6.8 (2H, m); 3.8 (3H, s);2.55 (3H, s) LCMS [M+H]=163

[0113] 100 g of BOC-protected indazole was dissolved in 600 mL of CCl₄,followed by addition of 1.1 equiv of NBS and 0.2 equiv of Bz2O. Reactionmix was vac-purged with argon and set to reflux for 5 h in presence oflight from a sun lamp. Reaction mixture was filtered over a pad of SGand concentrated. Residual oil was purified over a short SGC.Mono-bromide and mixed fractions of di-bromo derivative were obtained.

[0114] mono-bromide: 1H NMR (CDCL3): 7.7 (1H, d, 7.5 Hz); 7.6 (1H, bs);6.95 (1H, dd); 4.7 (2H, s); 3.9 (3H, s); 1.7 (9H, s);

[0115] di-bromide: 1H NMR (CDCL3): 8.05 (1H, d, J=7.5 Hz); 7.6 (1H, bs);7.0 (1H,dd); 6.85 (1H, s); 3.9 (3H, s); 1.7 (9H, s);

[0116] 78 g of 6-methoxy-3-methyl-1H-indazole was dissolved in 1L ofMeCN containing 111 equiv of tri-ethyl amine, 0.2 equiv of DMAP wascooled to −5° C.; followed by slow addition of Boc₂O (1.1 equiv) in 200mL of MeCN. After 2 h of stirring the reaction at room temperature thereaction mixture was evaporated to an oil which was partitioned betweenEtOAc and brine, dried over sodium sulphate and evaporated. The residuewas applied to a short SGC and eluted with 15% EtOAc in hexane.Evaporation gave Boc-protected product.

[0117] 1H NMR (CDCL3): 7.6 (1H, bs); 7.42 (1H, d, J=7.5 Hz); 6.85 (1H,dd); 3.8 (3H, s); 2.5 (3H, s); 1.7 (9H,s) LCMS [M+H]=263

[0118] To a solution of dibromide (23.2 g) in acetic acid was addedsodium acetate (22.5 g). The mixture was placed in oil bath and refluxedfor a couple of hours until reaction completed. The mixture was cooledto room temperature and then poured into ice/water to give desiredcompound as an off-white solid. The solid was isolated by filtration anddried over nitrogen atmosphere.

[0119]¹H NMR (CDCl₃): δ 10.23 (1H, s); 8.19 (1H, d); 7.02 (1H, dd); 6.96(1H, d); 3.90 (3H, s).

[0120] Step B:

[0121] To the intermediate from Step A was added triethyl orthoformate(40 ml) and heated to 130° C. for a couple of hours. The resultingmixture was concentrated to dry to give title compound as a brown solid.

[0122]¹H NMR (DMSO): δ 10.08 (1H, s); 7.98 (1H, d); 7.25 (1H, d); 7.02(1H, dd); 6.81 (1H, s); 3.82 (3H, s); 3.52 (4H, q); 1.11 (6H, t).

Preparative Example 6

[0123]

[0124] Oil free NaH (120 mg, 60% NaH in mineral oil was washed withhexanes 3 times.) suspended in DMF was added intermediate fromPreparative Example 2 (346 mg) at RT. After bubbles subsided, themixture was stirred at RT for 30 min and MOM-Cl (0.23 ml) was added.After the reaction completed, the mixture was poured into ice/water togive compound as a solid. The crude material was purified by silica gel(hexanes/ethyl acetate=3/1) to give title compound. ¹H NMR (CDCl₃): 7.72(1H, d); 7.06 (1H, dd); 6.99 (1H, d); 5.72 (2H, s); 3.94 (3H, s); 3.36(3H, s).

Preparative Example 7

[0125]

[0126] To a solution of intermediate from preparative Example 2 (1.00 g,5.75 mmol) dissolved in THF (15 mL) was added cyclopentyl magnesiumbromide (6.32 mL, 12.65 mmol) at 0° C. The reaction was allowed to warmto ambient temperature and was quenched with saturated NH₄Cl uponcompletion. The resulting reaction mixture was extracted with EtOAc andthe combined organic layers were washed with brine, dried over MgSO₄,and concentrated in vacuo. The product was purified via SiO₂ gelchromatography to yield 580 mg of the desired product. ¹H NMR (CDCl₃) δ:1.702 (2 H, m), 1.803 (2 H, m), 2.005 (4 H, m), 3.904 (3 H, s), 4.070 (1H, m), 6.915 (1 H, s), 7.010 (1 H, d), 8.272 (1 H, d).

Preparative Example 8

[0127]

[0128] The desired compound was prepared by a procedure similar to theone described for Preparative Example 7, but cyclohexyl magnesiumbromide was used in place of cyclopentyl magnesium bromide. ¹H NMR(CDCl₃) δ: 1.327 (1 H, m), 1.479 (2 H, m), 1.604 (2 H, m), 1.781 (1 H,m), 1.861 (2 H, m), 2.000 (2 H, m), 3.641 (1 H, m), 3.902 (3 H, s),6.923 (1 H, s), 7.008 (1 H, d), 8.259 (1 H, d).

EXAMPLE 1

[0129]

[0130] Indazole (0.55 mmoles from Preparative Example 3) startingmaterial obtained as above was dissolved in DMF (3 mL) followed by theaddition of sodium hydride (0.88 mmoles). The reaction was stirred atroom temperature for 15 min, followed by the addition of1-bromo-pinacolone (0.669 mmoles). The reaction was stirred at roomtemperature for 30 min. TLC and LC-MS analysis indicated completeconsumption of starting material concurrent with the formation of a newproduct spot. The reaction mixture was quenched by the addition ofwater. Standard aqueous work-up followed by purification of crude by SGCgave the desired product as white solid.

[0131] 1H NMR CDCL3: 8.3 (3H, m); 7.61 (1H, t, J=7.5 Hz); 7.52 (2H, dd,J=7.5 and 7.0 Hz); 7.04 (1H, dd, J=2 and 9 Hz); 6.56 (1H, d, J=2 Hz);5.4 (2H, s); 3.94 (3H, s); 1.4 (9H, s). LCMS [M+H]=351.

EXAMPLE 2

[0132]

[0133] Indazole (0.60 mmoles from Preparative Example 4) startingmaterial obtained as above was dissolved in DMF (3 mL) followed by theaddition of sodium hydride (0.88 mmoles). The reaction was stirred atroom temperature for 15 min, followed by the addition of1-bromo-pinacolone (0.669 mmoles). The reaction was stirred at roomtemperature for 30 min. TLC and LC-MS analysis indicated completeconsumption of starting material concurrent with the formation of a newproduct spot. The reaction mixture was quenched by the addition ofwater. Standard aqueous work-up followed by purification of crude by SGCgave the desired product as white solid.

[0134] 1H NMR in CDCL: 8.22 (1H, d, J=9 Hz); 6.97 (1H, dd, J=2 and 9Hz); 6.5 (1H, d J=2 Hz); 5.4 ( 2H, s); 3.94 (3H, s); 2.8 (1H, m); 1.38(9H, s); 1.27 (6H, d, J=6.5 Hz). LCMS=[M+H]=317

EXAMPLE 3

[0135]

[0136] 133 mg of indazole from Preparative Example 4 was dissolved indry DMF (3 mL), followed by the addition of sodium hydride (24.3 mg, 60%oil dispersion). After stirring at room temperature for 15 min. 0.2 mLof 2-ethyl-hexyl iodide was added. The reaction mixture was allowed tostirr for an additional 10 h. Upon standard aqueous work-up followed bypurification by SGC the desired product was obtained.

[0137] 1HN CDCL3: 8.22 (1H, d, J=8.5 Hz); 7.0 (1H, dd, J=8.5 and 2 Hz);6.75 (1H, d, J=2 Hz); 4.23 (2H, d, J=7.5 Hz); 3.9 (3H, s); 2.2 (1H, m);0.8-1.5 (15h, m). LCMS [M+H]=331

[0138] Examples 4 through 15 as shown below are made, with somemodification of the desired compound of Example 3, by alkylation of theindazole as described in Example 1. Additionally, analogs of Examples 1and 4-15 can be prepared following analogous procedures using theindazole of Preparative Example 4 or alternatively another indazoleprepared following procedures described herein.

EXAMPLE 4

[0139]

[0140] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 5.9 (1H, m); 5.15 (2H, m); 4.5(2H, t); 3.9 (3H, s); 2.8 (2H, m). LCMS [M+H]=307

EXAMPLE 5

[0141]

[0142] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.25 (2H, d, J=7.5 Hz); 3.9(3H, s); 1-2.2 (11H, m). LCMS [M+H]=349

EXAMPLE 6

[0143]

[0144] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.45 (2H, d, J=7.5 Hz); 3.9(3H, s); 3.0 (1H, m); 1.8-2.2 (6H, m). LCMS [M+H]=321

EXAMPLE 7

[0145]

[0146] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.35 (2H, d, J=7.5 Hz); 3.9(3H, s); 1.4 (1H, m); 0.7 (2H, m); 0.5 (2H, m). LCMS [M+H]=307

EXAMPLE 8

[0147]

[0148] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.2 (sH, s); 3.9 (3H, s); 1.1(9H, s). LCMS [M+H]=323

EXAMPLE 9

[0149]

[0150] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.25 (2H, d, J=7.5 hZ); 3.9(3H, s); 2.6 (1H, m); 1.02 (6H, d). LCMS [M+H]=309

EXAMPLE 10

[0151]

[0152] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.4 (2H, t, J=7.5 Hz); 3.9(3H, s); 2.0 (2H, m); 1.02 (3H, t, J=7.5 Hz). LCMS [M+H]=295

EXAMPLE 11

[0153]

[0154] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.4 (2H, t, J=7.5 Hz); 3.9(3H, s); 2.0 (2H, m); 0.8-1.5 (5H, m). LCMS [M+H]=337

EXAMPLE 12

[0155]

[0156] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 6.2 (1H, m); 5.0-5.4 (3H, m);3.9 (3H, s). LCMS [M+H]=293

EXAMPLE 13

[0157]

[0158] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.4 (2H, d, J=7.5 Hz); 3.9(3H, s); 2.1 (1H, m); 1.4 (4H, m); 1.0 (6H, t, J=7.5 Hz).

[0159] LCMS [M+H]=337

EXAMPLE 14

[0160]

[0161] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.4 (2H, t, J=7.5 Hz); 3.9(3H, s); 1.9 (2H, t, J=7.5 Hz); 1.1 (9H, s). LCMS [M+H]=337.

EXAMPLE 15

[0162]

[0163] 1H NMR CDCL3: 8.35 (3H, m); 7.6 (1H, t); 7.55 (2H, t); 7.1 (1H,dd, J=8.5 and 2 Hz); 6.8 (1H, d, J=2 Hz); 4.5 (2H, t, J=7.5 Hz); 3.9(3H, s); 1.9 (2H, m); 1.7 (1H, m); 1.05 (6H, d, J=7.5 Hz).

[0164] LCMS [M+H]=323

EXAMPLE 16

[0165]

[0166] Step A:

[0167] To a solution of 5-iodo-2-chloropyridine (2.56 g, 10.78 mmol) inTHF (10 mL) was added iPrMgBr dropwise at −78° C. The reaction stirredfor 1 h before Preparative Example 5 (1.71 g, 6.10 mmol) was added as asolution in THF (5 mL). After 2 h and the reaction was quenched with 1NNaOH and extracted with EtOAc. The combined organic layers were washedwith brine, dried over MgSO₄, and concentrated in vacuo. To a solutionof the crude product in toluene (50 mL) was added MnO₂ (2.173 g, 25.0mmol) and the reaction mixture was heated to 130° C. After 1 h thereaction was complete, filtered through a celite pad, and concentratedin vacuo. The crude product was dissolved in THF (10 mL) and 4 mL of 1NHCl was added dropwise. The reaction stirred at RT until TLC analysisindicated completion. The reaction mixture was cooled to 0° C. and thesolid precipitate was collected. ¹H NMR (CD₃OD) δ: 3.900 (3H, s), 7.013(1H, d), 7.062 (1H, s), 7.627 (1H, d), 8.672 (1H, d), 9.306 (1H, s).

[0168] Step B:

[0169] To a solution of the intermediate from Step A (1.00 g, 3.48 mmol)and Cs₂CO₃ (3.396 g, 10.45 mmol) in DMF (14 mL) was added1-chloropinacolone (0.681 mL, 5.22 mmol). After 40 min the reaction wascomplete and quenched with H₂O. The reaction mixture was extracted withEtOAc and the combined organic layers were washed with H₂O, brine, driedover MgSO₄, and concentrated in vacuo to yield the desired product. ¹HNMR (CD₃OD) δ: 1.344 (9H, s), 3.888 (3H, s), 6.947 (1H, s), 7.043 (1H,d), 7.625 (1H, d), 8.221 (1H, d), 8.624 (1H, d), 9.257 (1H, d).

[0170] Step C:

[0171] 40.6 mg (1.036 mmol) of NaH (60% dispersion in mineral oil) waswashed 3× with hexane and dried under nitrogen. Ethylene glycol (1 mL)was added to the dry NaH and the reaction stirred for 20 min at 60° C.To the reaction mixture was added the intermediate from Step B (100 mg,0.259 mmol) as a solution in TBF (1.5 mL). The reaction continued tostir overnight at 60° C. Upon completion, the TBF was removed in vacuo,diluted with EtOAc, washed with H₂O, brine, dried over MgSO₄, andconcentrated in vacuo. The crude residue was purified via silica gelchromatography.

[0172] 1.376 (9H, s), 3.889 (3H, s), 4.021 (2H, m), 4.608 (2H, m), 5.429(2H, s), 6.543 (1H, s), 6.223 (1H, d), 7.054 (1H, d), 8.336 (1H, d),8.541 (1H, d), 9.310 (1H, s).

EXAMPLE 17

[0173]

[0174] Step A:

[0175] To a solution of 5-bromo-2-methylpyridine (736 mg, 4.31 mmol) inTHF (15 mL) was added nBuLi dropwise (2.156 mL, 5.39 mmol, 2.5 M inhexanes) at −78° C. The reaction stirred for 1 h before PreparativeExample 5 (1.00 g, 3.59 mmol) was added as a solution in THF (5 mL). Thestarting material was consumed after 2 h and the reaction was quenchedwith 1N NaOH and extracted with EtOAc. The combined organic layers werewashed with brine, dried over MgSO₄, and concentrated in vacuo. Asolution of the crude product in toluene (20 mL) was added MnO₂ (0.414g, 4.77 mmol) and the reaction mixture was heated to 130° C. After 1 hthe reaction was complete, filtered through a celite pad, andconcentrated in vacuo. The crude product was dissolved in THF and 4 mLof 1N HCl was added dropwise. After 1 h reaction mixture was cooled to0° C. and the solid precipitate was collected. ¹H NMR (DMSO) δ: 2.553(3H, s), 3.832 (3H, s), 7.000 (1H, d), 7.089 (1H, s), 7.451 (1H, d),8.100 (1H, d), 8.430 (1H, d), 9.220 (1H, s).

[0176] Step B:

[0177] This compound was made as described in Step B of Example 16.

[0178]¹H NMR (CDCl₃) 8: 1.38 (9H, s), 2.65 (3H, s), 3.85 (3H, s), 5.22(2H, s), 6.56 (1H, s), 7.05 (1H, d), 7.32 (1H, d), 8.34 (1H, d), 8.45(1H, d), 9.50 (1H, s).

[0179] Step C:

[0180] To a stirring solution of the intermediate from Step B (74 mg,0.202 mmol) in CH₂Cl₂ was added MCPBA (67 mg, 0.303 mmol) at 0° C. TLCindicated the reaction was complete after 1.5 h and the reaction mixturewas concentrated in vacuo. The crude residue was dissolved in EtOAc andwashed with saturated sodium bisulfite, H₂O, brine, dried over MgSO₄,and concentrated in vacuo. Purified via silica gel chromatography. TheN-oxide was dissolved in CH₂Cl₂ and TFAA was added dropwise at 0° C.After 2 h the reaction was concentrated in vacuo and purified via silicagel chromatography.

[0181]¹H NMR (CDCl₃) δ: 1.373 (9H, s), 3.898 (3H, s), 4.882 (2H, s),5.428 (2H, s), 6.564 (1H, s), 7.066 (1H, d), 7.429 (1H, d), 8.352 (1H,d), 8.581 (1H, d), 9.541 (1H, s).

EXAMPLE 18

[0182]

[0183] Step A:

[0184] To a solution of 2-pyridineacetic acid, 5-bromo-α,α-difluoro-,ethyl ester (13.4 g; prepared according to “Ero, H.; Haneko, Y.;Sakamoto, T. Chem Pharm. Bull. 2000,48, 982.”) in ethanol was addedsodium borohydride (2.3 g) portion-wise at 0° C. After stirring at 0° C.for 1 hour, the mixture was poured into water and extracted with ethylacetate. The organic layer was washed with 1N NaOH_(aq), brine, dried(MgSO₄), and concentrated under reduced pressure to afford crudealcohol. The crude alcohol in methylene chloride was added imidazole(4.1 g) and TBS-Cl (8.3 g) at 0° C. The mixture was stirred for 1 hour.The reaction was poured into 0.1 N HCl_(aq) extracted with methylenechloride. The organic layer was washed with brine, dried (MgSO₄) andevaporated. The residue was purified by silica gel (100% methylenechloride) to give desired compound as a colorless oil.

[0185]¹H NMR (CDCl₃): δ 8.75 (1H, d); 7.95 (1H, dd); 7.57 (1H, d); 4.20(2H, t); 0.82 (9H, s); 0.02 (6H, s).

[0186] Step B:

[0187] The desired compound was prepared by a procedure similar to theone described for Example 16, Step A.

[0188]¹H NMR (DMSO): δ 9.35 (1H, d); 8.65 (1H, dd); 8.14 (1H, d); 7.88(1H, d); 7.10 (1H, d); 7.03 (1H, dd); 4.05 (2H, t); 3.85 (3H, s).

[0189] LC-MS (M+H)=334.2.

[0190] Step C:

[0191] The desired compound was prepared by a procedure similar to theone described for Example 16, Step B. This compound was purified bysilica gel (hexanes/ethyl acetate=1/1) and crystalized fromhexanes/ethyl acetate.

[0192]¹H NMR (CHCl₃): δ 9.53 (1H, d); 8.71 (1H, dd); 8.35 (1H, d); 7.88(1H, d); 7.08 (1H, dd); 6.57 (1H, d); 5.44 (2H, s); 4.32 (2H, t); 3.91(3H, s); 1.38 (9H, s).

[0193] LC-MS (M+H)=432.3.

EXAMPLE 19

[0194]

[0195] Step A:

[0196] To a solution of 2,5-dibromopyridine (2.4 g) in toluene was addedtributylallyltin (3.4 ml) and dichlorobis(triphenylphosphine) palladium(0.7 g) under nitrogen atmosphere. The mixture was refluxed for a coupleof hours and concentrated under reduced pressure. The residue wasre-dissolved in “wet ether” and added DBU (3 ml) slowly to give a cloudysolution. The mixture was filtered over a pad of silica gel andconcentrated. The residue was dissolved in methylenechloride/methanol=1/1 solution and cooled to −78° C. To this solutionwas bubbled though ozone until the reaction mixture became a blue color.The reaction was warmed to 0° C. and added sodium borohydride (0.5 g)portion-wise. After stirring at 0° C. for 1 hour, the mixture was pouredinto water and extracted with ethyl acetate. The organic layer waswashed with 1N NaOH_(aq), brine, dried (MgSO₄), and concentrated underreduced pressure to afford crude alcohol. The alcohol was purified bysilica gel (methylene chloride/ethyl acetate=1/1) to give desiredalcohol. To a solution of alcohol in methylene chloride was addedimidazole (0.4 g) and TBS-Cl (0.8 g) at 0° C. The mixture was stirredfor 1 hour. The reaction was poured into 0.1 N HCl_(aq) extracted withmethylene chloride. The organic layer was washed with brine, dried(MgSO₄) and evaporated. The residue was purified by silica gel (100%methylene chloride) to give desired compound.

[0197]¹H NMR (CDCl₃): δ 8.61 (1H, d); 7.73 (1H, dd); 7.14 (1H, d); 3.97(2H, t); 2.96 (2H, t); 0.86 (9H, s); −0.02 (6H, s).

[0198] Step B:

[0199] The desired compound was prepared by a procedure similar to theone described for Example 16, Steps A and B. This compound was purifiedby silica gel (hexanes/ethyl acetate=1/3).

[0200]¹H NMR (CHCl₃): δ 9.53 (1H, d); 8.54 (1H, dd); 8.35 (1H, d); 7.37(1H, d); 7.07 (1H, dd); 6.56 (1H, d); 5.45 (2H, s); 4.11 (2H, t); 3.90(3H, s); 3.18 (2H, t); 1.38 (9H, s).

[0201] LC-MS (M+H)=396.2.

EXAMPLE 20

[0202]

[0203] Step A:

[0204] To a solution of chloroiodopyridine (1.2 g), pyrrolidine (1.2 ml)and potassium carbonate (2.0 g) in DMF was heated 130° C. for 16 h. Themixture was cooled to RT and poured into ice/water to give crude solidmaterial. The title compound was crystallized from hexanes/ethyl acetate(0.73 g).

[0205]¹H NMR (CDCl₃): 8.30 (1H, d); 7.62 (1H, dd); 6.23 (1H, d); 3.43(4H, m); 2.03 (4H, m).

[0206] Step B:

[0207] To a solution of intermediate from Step A (274 mg) in THF wasadded isopropyl magnesium chloride (0.5 ml, 2N in diethyl ether) at −78°C. The mixture was warmed up to RT for a couple hours to completeiodide-magnesium exchange and re-cooled to −78° C. when PreparativeExample 5 (110 mg) was added to the reaction mixture. The resultingsolution was stirred at RT for 16 h and quenched with 1N NaOH, exactedwith EtOAc. The organic layer was washed with brine, dried overmagnesium sulfate and evaporated under vacuum. The residue was dissolvedin 96% formic acid and heated to 100° C. for 16 h. After cooled to RT,the mixture was diluted with 1N NaOH to pH=9. The mixture was exactedwith EtOAc, brine, dried over magnesium sulfate and evaporated undervaccum to give crude material.

[0208] Step C:

[0209] The title compound was prepared as described in PreparativeExample 6, using chloropinacolone instead of MOM-Cl. The final compoundwas purified by silica gel (hexanes/ethyl acetate=1/1). ¹H NMR (CDCl₃):9.46 (1H, d); 8.40 (1H, d); 8.32 (1H, d); 7.00 (1H, dd); 6.53 ( 1H, d);6.48 (1H, d); 5.43 (2H, s); 3.89 (3H, s); 3.63 (4H, br. s); 2.08 (4H,br.s); 1.37 (9H, s). LCMS (M+H)=421.4.

EXAMPLE 21

[0210]

[0211] Step A: The title compound was prepared by a procedure similar tothe one described for Example 20, Step A. The reaction useddimethylamine hydrogen chloride and potassium carbonate instead ofpyrrolidine.

[0212]¹H NMR (CDCl₃): 8.31 (1H, d); 7.64 (1H, dd); 6.37 (1H, d); 3.08(6H,s).

[0213] Step B:

[0214] The title compound was prepared by a procedure similar to the onedescribed for Example 20, Step B and C by using intermediate fromExample 21, Step A instead of Example 20, Step A. ¹H NMR (CDCl₃): 9.41(1H, d); 8.41 (1H, dd); 8.32 (1H, d); 7.01 (1H, dd); 6.60 ( 1H, d); 6.53(1H, d); 5.42 (2H, s); 3.89 (3H, s); 3.24 (6H, s); 1.37 (9H, s).

[0215] LCMS (M+H)=395.4.

EXAMPLE 22

[0216]

[0217] To a solution of intermediate from Example 21, Step A in THF wasadded isopropyl magnesium chloride (2N in diethyl ether) at −78° C. Themixture was warmed up to RT for a couple hours to completeiodide-magnesium exchange and re-cooled to −78° C. when PreparativeExample 6 (110 mg) was added to the reaction mixture. The resultingsolution was stirred at RT for 16 h and quenched with 1N NaOH, exactedwith EtOAc. The organic layer was washed with brine, dried overmagnesium sulfate and evaporated under vacuum. The title compound waspurified by silica gel (methylene chloride/ethyl acetate=10/1).

[0218]¹H NMR (CDCl₃): 9.46 (1H, d); 8.45 (1H, dd); 8.31 (1H, d); 7.04(1H, dd); 6.98 ( 1H, d); 6.62 (1H, d); 5.77 (2H, s); 3.94 (3H, s); 3.39(3H, s); 3.25 (4H, s);

[0219] LCMS (M+H)=342.2.

EXAMPLE 23

[0220]

[0221] Step A:

[0222] To a solution of 2-chloro-5-iodopyridine in THF was addedisopropyl magnesium chloride (0.5 ml, 2N in diethyl ether) at −78° C.The mixture was warmed up to RT for a couple hours to completeiodide-magnesium exchange and re-cooled to −78° C. when Intermediatefrom Preparative Example 2 was added to the reaction mixture. Theresulting solution was stirred at RT for 16 h and quenched with 1N NaOH,exacted with EtOAc. The organic layer was washed with brine, dried overmagnesium sulfate and evaporated under vacuum.

[0223] Step B:

[0224] The title compound was prepared as described in PreparativeExample 6, using chloropinacolone instead of MOM-Cl. The final compoundwas purified by silica gel (hexanes/ethyl acetate=3/1).

[0225]¹H NMR (CDCl₃): 9.41 (1H, d); 8.53 (1H, dd); 8.33 (1H, d); 7.49(1H, dd); 7.07 ( 1H, dd); 6.58 (1H, d); 5.43 (2H, s); 3.91 (3H, s); 1.37(9H, s).

[0226] LCMS (M+H)=386.3.

EXAMPLE 24

[0227]

[0228] To a solution of intermediate from Example 23 in THF was addedsodium methoxide (3 eq. 25% in methanol). The mixture was refluxed untilreaction completed and quenched with 1N HCl. The mixture was extractedwith ethyl acetate, brine, dried over magnesium sulfate and evaporatedunder vacuum. The residue was purified with silica gel (hexanes/ethylacetate=2/1).

[0229]¹H NMR (CDCl₃): 9.35 (1H, d); 8.50 (1H, dd); 8.33 (1H, d); 7.04(1H, dd); 6.86 ( 1H, d); 6.55 (1H, d); 5.43 (2H, s); 4.06 (3H, s); 3.90(3H, s); 1.38 (9H, s).

[0230] LCMS (M+H)=386.3.

EXAMPLE 25

[0231]

[0232] To 195 mg of NaH (60% dispersion in oil washed with hexane) wasadded DMF (10 mL) and Preparative Example 7 (597 mg, 2.44 mmol). Thereaction stirred at room temperature for 30 min before1-chloropinacolone (3.81 mL, 2.92 mmol) was added. After 20 min thereaction was quenched with H₂O and diluted with EtOAc. The aqueous layerwas extracted with EtOAc and the combined organic layers were washedwith H₂O, brine, dried over MgSO₄, and concentrated in vacuo. The crudematerial was purified via silica gel chromatography to yield.

[0233]¹H NMR (CDCl₃) δ: 1.361 (9 H, s), 1.683 (2 H, m), 1.788 (2 H, m),1.974 (4 H, m), 3.872 (3 H, s), 4.029 (1 H, m), 5.372 (2 H, s), 6.514 (1H, s), 6.986 (1 H, d), 8.267 (1 H, d).

EXAMPLE 26

[0234]

[0235] Using Preparative Example 8, this compound was prepared asdescribed in Example 25. The title compound was purified via SiO₂preparatory plate chromatography. ¹H NMR (CDCl₃) δ: 1.285-1.575 (15 H,m), 1.833 (2 H, d), 1.994 (2 H, d), 3.615 (1 H, m), 3.860 (3 H, s),5.372 (2 H, s), 6.490 (1 H, s), 6.981 (1 H, d), 8.254 (1 H, d).

EXAMPLE 27

[0236]

[0237] Using the intermediate from Example 23, this compound wasprepared as described in Example 16, Step C but 2-methoxy-ethanol wasused in place of ethylene glycol. The title compound was purified viaSiO₂ preparatory plate chromatography. ¹H NMR (CDCl₃) δ: 1.371 (9 H, s),3.475 (3 H, s), 3.802 (2 H, t), 3.889 (3H, s), 4.620 (2H, t), 5.424 (2H,s), 6.542 (1H, s), 6.923 (1H, d), 7.043 (1H, d), 8.337 (1H, d), 8.501(1H, d), 9.299 (1H, s).

EXAMPLE 28

[0238]

[0239] di-tert-butyl4-{[1-(3,3-dimethyl-2-oxobutyl)-6-methoxy-1-H-indazole-3-yl]carbonyl}hydroxylethyl benzyl

[0240] Step 1

[0241] 100 g of fluoro-acetophenone in 400 mL of ethylene glycol wasstirred at room temperature with hydrazine (0.624 mol, 20 g) for 4hafter which the reaction mixture was heated to 150 C for 48 h. TLCanalysis indicated complete reaction. Partitioned the reaction mixtureinto dichloromethane and brine. Dried organic phase over sodium sulphateand evaporated to a solid. Re-crystallized from hexane/dicholomethanegave indazole.

[0242] 1H NMR (CDCL3): 7.5 (1H, d, 7.5 Hz); 6.8 (2H, m); 3.8 (3H, s);2.55 (3H, s) LCMS [M+H]=163

[0243] Step 2

[0244] 78 g of indazole was dissolved in 1L of MeCN containing 1.1 equivof tri-ethyl amine, 0.2 equiv of DMAP was cooled to −5 C; followed byslow addition of Boc2O (1.1 equiv) in 200 mL of MeCN. After 2 h ofstirring the reaction at rt the reaction mixture was evaporated to anoil which was partitioned between EtOAc and brine, dried over sodiumsulphate and evaporated. The residue was applied to a short SGC andeluted with 15% EtOAc in hexane. Evaporation gave product.

[0245] 1H NMR (CDCL3): 7.6 (1H, bs); 7.42 (1H, d, J=7.5 Hz); 6.85 (1H,dd); 3.8 (3H, s); 2.5 (3H, s); 1.7 (9H, s)

[0246] LCMS [M+H]=263

[0247] Step 3

[0248] 100 g of indazole was dissolved in 600 mL of CCl₄, followed byaddition of 1.1 equiv of NBS and 0.2 equiv of Bz2O. Reaction mix wasvac-purged with argon and set to reflux for 5 h in presence of lightfrom a sun lamp. Reaction mixture was filtered over a pad of SG andconcentrated. Residual oil was purified over a short SGC. 85 g of purebromide was obtained. Mixed fractions yielded di-bromo derivative

[0249] mono-bromide: 1H NMR (CDCL3): 7.7 (1H, d, 7.5 Hz); 7.6 (1H, bs);6.95 (1H, dd); 4.7 (2H, s); 3.9 (3H, s); 1.7 (9H, s);

[0250] di-bromide: 1H NMR (CDCL3): 8.05 (1H, d, J=7.5 Hz); 7.6 (1H, bs);7.0 (1H,dd); 6.85 (1H, s); 3.9 (3H, s); 1.7 (9H, s);

[0251] Step 4

[0252] 5 g of bromide was dissolved in 10 mL of DMSO, cooled to 0 Cfollowed by addition of 2.5 equiv of TMANO (trimethyl amine N-oxide).Reaction was stirred for 0.5 h then a standard work-up and SG padfilteration gave desired product quantitatively. LCMS [M+H]=277

[0253] 1H NMR (CDCL3): 10.2 (1H, s); 8.1 (1H, d, J=7.5 Hz); 7.6 (1H,bs); 7.0 (1H, dd); 3.9 (3H, s); 1.7 (9H, s);

[0254] Step 5

[0255] Glasswares were flame dried under high vacuum

[0256] To neat iodo-benzyl alcohol derivative (3.6 g, 10 mmol) in theflask was slowly added isopropyl MgCl ( 5 mL, 2M solution). Afterstirring at rt for 2 hr, indazole derivative (1.1 g, 4 mmol) in 15 mLTHF was added. The reaction mixture was stirred at rt for 2 hr. LC-MSshowed the reaction was complete. Pour the reaction mixture into 30 mLsaturated NH4Cl, followed by adding 40 mL ether. The organic layer wasseparated, the aqueous layer was extracted by ether (40 mL). Thecombined organic layers were washed with saturated K2CO3 (2×30 mL),water (40 mL) and brine (20 mL). The solvent was removed, the residuewas used for next step reaction without further purification. LCMS[M+H]=499

[0257] Steps 6 and 7

[0258] To a solution of indazole (crude from step 5) in 20 mLdichloromethane was added 5 g celite and 4.3 g of PCC (MW 215.56, ˜2eq). The reaction mixture was stirred at rt for 2 hr. LC-MS showed thereaction was completed LCMS [M+H]=497. The reaction mixture wasfiltered. The solvent was removed, the residue was dissolved in 10 mLMeOH, and added 20 mL 2N HCl. After stirring for 1 hr at rt LCMS and TLCanalysis indicated complete reaction. The reaction mixture was extractedwith EtOAc (2×30 mL). The solvent was removed, the residue was used fornext step reaction without further purification. LCMS [M+H]=283

[0259] Step 8.

[0260] To a solution of indazole (342 mg crude prod from step 7, ˜10mmol)) in 15 mL acetone was added 1.5 g of K2CO₃ and 1.5 mLBromopinacolone (Mw179.06, d1.326, 2.0 g, 11 mmol). The reaction mixturewas stirred at 80° C. in a seal tube for 2 hr. After filtered off salts,the solvent was removed, the residue was purified by HPFC to give whitesolid product.

[0261] 1H NMR (CDCL3)=8.3 (3H, m); 7.5 (1H, d, J=7.5 Hz); 7.05 (1H, dd);7.6 (1H, bs); 5.4 (2H, s); 4.8 (2H, bs); 3.9 (3H, s); 1.38 (9H, s)

[0262] LCMS [M+H]=381

[0263] Functional Assays

[0264] A. Maxi-K Channel

[0265] The identification of inhibitors of the Maxi-K channel can beaccomplished using Aurora Biosciences technology, and is based on theability of expressed Maxi-K channels to set cellular resting potentialafter transient transfection of both α and β subunits of the channel inTsA-201 cells. In the absence of inhibitors, cells display ahyperpolarized membrane potential, negative inside, close to E_(K) (−80mV) which is a consequence of the activity of the Maxi-K channel.Blockade of the Maxi-K channel will cause cell depolarization. Changesin membrane potential can be determined with voltage-sensitivefluorescence resonance energy transfer (FRET) dye pairs that use twocomponents, a donor coumarin (CC₂DMPE) and an acceptor oxanol(DiSBAC₂(3)). Oxanol is a lipophilic anion and distributes across themembrane according to membrane potential. Under normal conditions, whenthe inside of the cell is negative with respect to the outside, oxanolis accumulated at the outer leaflet of the membrane and excitation ofcoumarin will cause FRET to occur. Conditions that lead to membranedepolarization will cause the oxanol to redistribute to the inside ofthe cell, and, as a consequence, to a decrease in FRET. Thus, the ratiochange (donor/acceptor) increases after membrane depolarization.

[0266] Transient transfection of the Maxi-K channel in TsA-201 cells canbe carried out as previously described (Hanner et al. (1998) J. Biol.Chem. 273, 16289-16296) using FUGENE6™ as the transfection reagent.Twenty four hours after transfection, cells are collected inCa²⁺-Mg²⁺-free Dulbecco's phosphate-buffered saline (D-PBS), subjectedto centrifugation, plated onto 96-well poly-d-lysine coated plates at adensity of 60,000 cells/well, and incubated overnight. The cells arethen washed 1× with D-PBS, and loaded with 100 μl of 4 μM CC₂DMPE-0.02%pluronic-127 in D-PBS. Cells are incubated at room temperature for 30min in the dark. Afterwards, cells are washed 2× with D-PBS and loadedwith 100 μl of 6 μM DiSBAC₂(3) in (mM): 140 NaCl, 0.1 KCl, 2 CaCl₂, 1MgCl₂, 20 Hepes-NaOH, pH 7.4, 10 glucose. Test compounds are dilutedinto this solution, and added at the same time. Cells are incubated atroom temperature for 30 min in the dark.

[0267] Plates are loaded into a voltage/ion probe reader (VIPR)instrument, and the fluorescence emission of both CC₂DMPE and DiSBAC₂(3)are recorded for 10 sec. At this point, 100 μl of high-potassiumsolution (mM): 140 KCl, 2 CaCl₂, 1 MgCl₂, 20 Hepes-KOH, pH 7.4, 10glucose are added and the fluorescence emission of both dyes recordedfor an additional 10 sec. The ratio CC₂DMPE/DiSBAC₂(3), before additionof high-potassium solution equals 1. In the absence of any inhibitor,the ratio after addition of high-potassium solution varies between1.65-2.0. When the Maxi-K channel has been completely inhibited byeither a known standard or test compound, this ratio remains at 1. It ispossible, therefore, to titrate the activity of a Maxi-K channelinhibitor by monitoring the concentration-dependent change in thefluorescence ratio.

[0268] The compounds of this invention were found to causeconcentration-dependent inhibition of the fluorescence ratio with IC₅₀'sin the range of about 1 nM to about 20 μM, more preferably from about 10nM to about 500 nM.

[0269] B. Electrophysiological Assays of Compound Effects onHigh-Conductance Calcium-activated Potassium Channels

[0270] Human Non-Pigmented Ciliary Epithelial Cells

[0271] The activity of high-conductance calcium-activated potassium(maxi-K) channels in human non-pigmented ciliary epithelial cells wasdetermined using electrophysiological methods. Currents through maxi-Kchannels were recorded in the inside-out configuration of the patchclamp technique, where the pipette solution faces the extracellular sideof the channel and the bath solution faces the intracellular side.Excised patches contained one to about fifty maxi-K channels. Maxi-Kchannels were identified by their large single channel conductance(250-300 pS), and by sensitivity of channel gating to membrane potentialand intracellular calcium concentration. Membrane currents were recordedusing standard electrophysiological techniques. Glass pipettes (Garner7052) were pulled in two stages with a Kopf puller (model 750), andelectrode resistance was 1-3 megohms when filled with saline. Membranecurrents were recorded with EPC₉ (HEKA Instruments) or Axopatch 1D (AxonInstruments) amplifiers, and digital conversion was done with ITC-16interfaces (Instrutech Corp). Pipettes were filled with (mM); 150 KCl,10 Hepes, 1 MgCl₂, 0.01 CaCl₂, 3.65 KOH, pH 7.20. The bath(intracellular) solution was identical, except, in some cases, calciumwas removed, 1 mM EGTA was added and 20 mM KCl was replaced with 20 mMKF to eliminate calcium to test for calcium sensitivity of channelgating. Drugs were applied to the intracellular side of the channel bybath perfusion.

[0272] Human non-pigmented ciliary epithelial cells were grown in tissueculture as described (Martin-Vasallo, P., Ghosh, S., and Coca-Prados,M., 1989, J. Cell. Physiol. 141, 243-252), and plated onto glass coverslips prior to use. High resistance seals (>1 Gohm) were formed betweenthe pipette and cell surface, and inside out patches were excised.Maxi-K channels in the patch were identified by their gating properties;channel open probability increased in response to membranedepolarization and elevated intracellular calcium. In patches used forpharmacological analysis, removing intracellular calcium eliminatedvoltage-gated currents. Maxi-K currents were measured after depolarizingvoltage steps or ramps that caused channel opening.

[0273] The compounds of this invention were applied to the intracellularside of the channel in appropriate concentrations (0.001 to 100 μM). Thecompounds reduced channel open probability, and this effect was reversedupon washout of compounds from the experimental chamber. The IC50 forblock of maxi-K channels under these conditions for the compounds ofthis invention ranged from about 0.5 nM to about 10 μM.

What is claimed is:
 1. A compound of the structural formula I:

or a pharmaceutically acceptable salt, enantiomer, diastereomer ormixture thereof: wherein, R represents hydrogen, or C₁₋₆ alkyl; Xrepresents —(CHR₇)_(p)—, —(CHR₇)_(p)CO—; Y represents —CO(CH₂)_(n)—,CH_(2,) or —CH(OR)—; Q represents CR^(y); R^(y) represents H, or C₁₋₆alkyl; R_(w) represents H, C₁₋₆ alkyl, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆alkyl, —SO₂N(R)₂, —SO₂C₁₋₆ alkyl, —SO₂C₆₋₁₀ aryl, NO₂, CN or —C(O)N(R)₂;R₂ represents hydrogen, C₁₋₁₀ alkyl, OH, C₂₋₆ alkenyl, C₁₋₆ alkylSR,—(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆ alkoxy, —(CH₂)_(n)C₃₋₈cycloalkyl, —(CH₂)_(n)C₃₋₁₀ heterocyclyl, —N(R)₂, —COOR, or—(CH₂)_(n)C₆₋₁₀ aryl, said alkyl, heterocyclyl, or aryl optionallysubstituted with 1-3 groups selected from R^(a); R₃ represents hydrogen,C₁₋₁₀ alkyl, —(CH₂)_(n)C₃₋₈ cycloalkyl, —(CH₂)_(n)C₃₋₁₀ heterocyclyl,—(CH₂)_(n)COOR, —(CH₂)_(n)C₆₋₁₀ aryl, —(CH₂)_(n)NHR₈, —(CH₂)_(n)N(R)₂,—(CH₂)_(n)N(R₈)₂, —(CH₂)_(n)NHCOOR, —(CH₂)_(n)N(R₈)CO₂R,—(CH₂)_(n)N(R₈)COR, —(CH₂)_(n)NHCOR, —(CH₂)_(n)CONH(R₈), aryl,—(CH₂)_(n)C₁₋₆ alkoxy, CF₃, —(CH₂)_(n)SO₂R, —(CH₂)_(n)SO₂N(R)₂,—(CH₂)_(n)CON(R)₂, —(CH₂)_(n)CONHC(R)₃, —(CH₂)_(n)CONHC(R)₂CO₂R,—(CH₂)_(n)COR₈, nitro, cyano or halogen, said alkyl, alkoxy,heterocyclyl, or aryl optionally substituted with 1-3 groups of R^(a);or R₂ and R₃ taken together with the intervening Q form a 3-10 memberedcarbocyclic or heterocyclic carbon ring optionally interrupted by 1-2atoms of O, S, C(O) or NR, and optionally having 1-4 double bonds, andoptionally substituted by 1-3 groups selected from R^(a); or R₂ and R₃taken together with the intervening Q represent OR; R₄ and R₅independently represent hydrogen, C₁₋₆ alkoxy, OH, C₁₋₆ alkyl, COOR,SO_(q)C₁₋₆ alkyl, COC₁₋₆ alkyl, SO₃H, —O(CH₂)_(n)N(R)₂, —O(CH₂)_(n)CO₂R,—OPO(OH)₂, CF₃, OCF₃—N(R)₂, nitro, cyano, C₁₋₆ alkylamino, or halogen;and R₆ represents hydrogen, C₁₋₁₀ alkyl, —(CH₂)_(n)C₆₋₁₀ aryl,NR_(c)R_(d), —NR(CH₂)_(n)C₆₋₁₀ aryl, —N((CH₂)_(n)C₆₋₁₀ aryl)₂,—(CH₂)_(n)C₃₋₁₀ heterocyclyl, —NR(CH₂)_(n)C₃₋₁₀ heterocyclyl,—N((CH₂)_(n)C₃₋₁₀ heterocyclyl)₂ (C₆₋₁₀ aryl)O—, —(CH₂)_(n)C₃₋₈cycloalkyl, —COOR, —C(O)CO₂R, said aryl, heterocyclyl and alkyloptionally substituted with 1-3 groups selected from R^(a), wherein theR^(a)(s) can be attached to any carbon atom or heteroatom selected fromN and S; R_(c) and R_(d) independently represent H, C₁₋₆ alkyl, C₂₋₆alkenyl, C₁₋₆ alkylSR, —(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆ alkoxy, or—(CH₂)_(n)C₃₋₈ cycloalkyl; or R_(c) and R_(d) taken together with theintervening N atom form a 4-10 membered heterocyclic carbon ringoptionally interrupted by 1-2 atoms of O, S, C(O) or NR, and optionallyhaving 1-4 double bonds, and optionally substituted by 1-3 groupsselected from R^(a); R₇ represents hydrogen, C₁₋₆ alkyl, —(CH₂)_(n)COORor —(CH₂)_(n)N(R)₂, R₈ represents —(CH₂)_(n)C₃₋₈ cycloalkyl,—(CH₂)_(n 3-10) heterocyclyl, C₁₋₆ alkoxy or —(CH₂)_(n)C₅₋₁₀ heteroaryl,—(CH₂)_(n)C₆₋₁₀ aryl said heterocyclyl, aryl or heteroaryl optionallysubstituted with 1-3 groups selected from R^(a); R^(a) represents F, Cl,Br, I, CF₃, N(R)₂, NO₂, CN, —O—, —COR₈, —CONHR₈, —CON(R₈)₂,—O(CH₂)_(n)COOR, —NH(CH₂)_(n)OR, —COOR, —OCF₃, CF₂CH₂OR, —NHCOR, —SO₂R,—SO₂NR₂, —SR, (C₁-C₆ alkyl)O—, —(CH₂)_(n)O(CH₂)_(m)OR, —(CH₂)_(n)C₁₋₆alkoxy, (aryl)O—, —(CH₂)_(n)OH, (C₁-C₆ alkyl)S(O)M—, H₂N—C(NH)—, (C₁-C₆alkyl)C(O)—, (C₁-C₆ alkyl)OC(O)NH—, —(C₁-C₆ alkyl)NR_(w)(CH₂)_(n)C₃₋₁₀heterocyclyl-R_(w), —(C₁-C₆ alkyl)O(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w),—(C₁-C₆ alkyl)S(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₁-C₆ alkyl)—C₃₋₁₀heterocyclyl-R_(w), —(CH₂)_(n)—Z¹—C(═Z²)N(R)₂, —(C₂₋₆alkenyl)NR_(w)(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆alkenyl)O(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆alkenyl)S(CH₂)_(n)C₃₋₁₀ heterocyclyl-R_(w), —(C₂₋₆ alkenyl)-C₃₋₁₀heterocyclyl-R_(w), —(C₂₋₆ alkenyl)—Z¹—C(═Z²)N(R)₂, —(CH₂)_(n)SO₂R,—(CH₂)_(n)SO₃H, —(CH₂)_(n)PO(OR)₂, C₃₋₁₀cycloalkyl, C₆₋₁₀ aryl, C₃₋₁₀heterocyclyl, C₂₋₆ alkenyl, and C₁-C₁₀ alkyl, said alkyl, alkenyl,alkoxy, heterocyclyl and aryl optionally substituted with 1-3 groupsselected from C₁-C₆ alkyl, halogen, (CH₂)_(n)OH, CN, NO₂, CON(R)₂ andCOOR; Z¹ and Z² independently represents NR_(w), O, CH₂, or S; m is 0-3;n is 0-3; p is 0-3 and q is 0-2.
 2. A compound according to claim 1wherein R₆ is C₁₋₁₀ alkyl, (CH₂)_(n)C₆₋₁₀ aryl, (CH₂)_(n)C₃₋₁₀heterocyclyl, NR_(c)R_(d) or (CH₂)_(n)C₃₋₈ cycloalkyl, said aryl,heterocyclyl and alkyl optionally substituted with 1 to 3 groups ofR^(a).
 3. A compound according to claim 1 wherein Y is —CO(CH₂)_(n), nis O, R₂ is Cl-lo alkyl or C₁₆ alkylOR and R₃ is C₁₋₁₀ alkyl,(CH₂)_(n)C₃₋₁₀ heterocyclyl, X is (CHR₇)_(p)CO—, and p is 1-3 saidheterocyclyl and alkyl optionally substituted with 1 to 3 groups ofR^(a).
 4. A compound which is: TABLE 1

R1 R2

TABLE 2

wherein R₁ is

and X is N or CH;

or a pharmaceutically acceptable salt, enantiomer, diastereomer ormixture thereof.
 5. A method for treating ocular hypertension orglaucoma comprising administration to a patient in need of suchtreatment a therapeutically effective amount of a compound of structuralformula I of claim
 1. 6. A method for treating macular edema, maculardegeneration, increasing retinal and optic nerve head blood velocity,increasing retinal and optic nerve oxygen tension, and/or aneuroprotective effect comprising administration to a patient in need ofsuch treatment a pharmaceutically effective amount of a compound ofclaim 1; or a pharmaceutically acceptable salt, enantiomer, diastereomeror mixture thereof.
 7. A method of preventing repolarization orhyperpolarization of a mammalian cell containing potassium channel or amethod of treating Alzheimer's Disease, depression, cognitive disorders,and/or arrhythmia disorders in a patient in need thereof comprisingadministering a pharmaceutically effective amount of a compoundaccording to claim 1, or a pharmaceutically acceptable salt, enantiomer,diastereomer or mixture thereof.
 8. A method of treating diabetes in apatient in need thereof comprising administering a pharmaceuticallyeffective amount of a compound according to claim 1, or apharmaceutically acceptable salt, enantiomer, diastereomer or mixturethereof.
 9. A composition comprising a compound of formula I of claim 1and a pharmaceutically acceptable carrier.
 10. The composition accordingto claim 9 wherein the compound of formula I is applied as a topicalformulation, said topical formulation administered as a solution orsuspension and optionally containing xanthan gum or gellan gum.
 11. Acomposition according to claim 9 wherein an active ingredient belongingto the group consisting of: β-adrenergic blocking agent,parasympatho-mimetic agent, sympathomimetic agent, carbonic anhydraseinhibitor, EP4 agonist, a prostaglandin or derivative thereof,hypotensive lipid, neuroprotectant, and/or 5-HT2 receptor agonist isoptionally added.
 12. A composition according to claim 11 wherein theβ-adrenergic blocking agent is timolol, betaxolol, levobetaxolol,carteolol, or levobunolol; the parasympathomimetic agent is pilocarpine;the sympathomimetic agent is epinephrine, brimonidine, iopidine,clonidine, or para-aminoclonidine, the carbonic anhydrase inhibitor isdorzolamide, acetazolamide, metazolamide or brinzolamide; theprostaglandin is latanoprost, travaprost, unoprostone, rescula, orS1033, the hypotensive lipid is lumigan, the neuroprotectant iseliprodil, R-eliprodil or memantine; and the 5-HT2 receptor agonist is1-(2-aminopropyl)-3-methyl-1H-imdazol-6-ol fumarate or2-(3-chloro-6-methoxy-indazol-1-yl)-1-methyl-ethylamine.